JP2002526444A - Methods and compositions for treating receptor-mediated diseases - Google Patents

Abstract

  (57) [Summary] Disclosed are small molecule agents for treating sulfate receptor-mediated diseases.

Description

DETAILED DESCRIPTION OF THE INVENTION

BACKGROUND OF THE INVENTION Receptors on the cell surface play a major role in the initial infection of pathogenic viruses, bacteria and fungi. The host specificity of many pathogens is determined when they first contact and attach to eukaryotic plant or animal cells. For example, some fungal pathogens to plants release a host-selective (or host-specific) toxin (HST) as a host-recognizing factor when spores germinate at the site of infection of the plant. This released toxin specifically binds to a single receptor on the host cell and initiates a signaling mechanism to exert a pleiotropic effect on the cell before the pathogen enters the host (Kohmoto and Otani (1991) Experientia 47: 755-64).
The most important of these are those that ascribe the general and inducible protective response of the cell. This is done by a signal from the HST, which is introduced in a path close to or near the step of cell membrane modulation caused directly or indirectly by the HST. Thus, the fungal spores can invade so-called "drug-killed cells" and complete the first colony formation with this host. Such host recognition processes may not necessarily require the death of host cells, even in the case of biocidal parasites. At the molecular level, strict stereochemical precision, such as keys and keyholes, between host receptors and fungal pathogens is required for pathogen recognition through HST-mediated recognition. The importance of the pathogen / plant host receptor interaction is
In addition, genetic studies on plant disease resistance have provided evidence. For example, in plants, a major locus of lettuce pathogen resistance has been isolated and found to encode a family of receptor-like genes (Meyers et al. (1998) Plant
Cell 10: 1833-46).

[0002] Infection of animal cells with pathogens likewise depends on the initial interaction between bacteria and proteins on the surface of host cells. Enterotoxigenic Escherichia coli (ETEC) causes intestinal diarrhea in animals and humans, but first infects host organisms by first attaching to receptors on mucosal surfaces (Mour).
icout (1991) Eur J Epidemiol 7: 588-604). For ETEC infections,
In the process of bacteria recognizing and attaching a host receptor, a function of attaching a specific bacterial pilus, which acts as a lectin in the process of attachment, is required. It has been shown that the ability of various pathogens to attach to and form colonies in the host cell layer involves the production of such hair-like ciliated adherents or pili, protruding from the surface of bacterial cells. Have been. The production of these pili is controlled by a specific plasmid in the pathogen, and if this plasmid is lost, the bacterium becomes nontoxic even if it can continue to produce toxins (Shaeffer (1993) Urologe A 32: 7-15). Thus, attachment of bacterial pili to host cell surface receptors plays an important role in the development of a number of infectious diseases, including intestinal E. coli infections, ureteral infections, and rheumatic fever.

In addition, there are many examples of receptor-mediated infection of human host cells by pathogenic viruses. Measles virus infects human host cells via a CD46 cell surface receptor (Gerlier et al. (1995) Treands Microbiol 3: 338-45), and poliovirus similarly binds to specific human poliovirus receptors. The effect is important for viral RNA uncoating and subsequent infectious events (Racaniello (1996) Proc Natl Acad Sci USA 93: 11378-81), and for HIV gp120.
Upon binding of the viral envelope glycoprotein and subsequent infection of T cells, CD4
The role played by cell surface receptors is well documented (eg, Bour et al. (1
995) Microbiol Rev 59: 63-93).

[0004] Interestingly, diseases without the involvement of pathogens, such as viruses, bacteria or fungi, may also involve host cell surface receptors in the onset or progression of the disease. An interesting case in this regard is the development of infectious spongiform encephalopathy in humans and animals, in which host receptors purposely promote the transfer of infectious prion protein from cell to cell within the infected host. (Martins (1999) Braz J Med Biol Res 32: 853-9). Another example is certain types of cancer. Thus, chemical compounds that can attach to cell surface receptors would be useful in treating and preventing a wide range of infectious and non-infectious diseases and conditions.

[0005] Lectins are a class of proteins known in nature to bind to a wide variety of cell surface moieties. Lectins are a family of animal and plant cell proteins that bind with high affinity to specific plant and animal cell surface oligosaccharides. For example, Concavillin A is a plant protein that binds to internal and non-reducing terminal α-mannosyl residues, while wheat malt agglutinin has terminal N-
It is a plant protein that binds to acetylglucosamine residues. Interestingly, the binding of such plant lectins to the outer surface of eukaryotic cells can have significant biological consequences on the bound cells. For example, binding of Concavillin A stimulates mitosis, and the lectin wheat phytohemagglutin causes aggregation of mammalian erythrocytes. Plant and microbial lectins have also been shown to affect a number of other biological processes (Rudiger (199
8) Additional tests at Acta Anat (Basel) 161: 130-52). For example, lectins in the roots of cereals specifically recognize the N-acetyl-D-glucosamine extracellular polysaccharide component of azospirillum, a bacterium of the genus Rhizobium, thereby mediating a symbiotic relationship between legume and rhizobia ( Skvortsov and Ignatov (1998) FEMS Microbiol Lett 165: 223-9)
. Similarly, lectins have been implicated in the recognition of plant hosts by phytopathogenic fungi (Hardham and Mitchell (1998) Fungal Genet Biol 24: 252-84).

[0006] Many more animal lectins have been discovered and implicated in a wide range of biological functions. For example, selectins mediate specific cell-to-cell adhesion events, such as directing lymphocyte trafficking by binding lymphocytes to activated endothelial cells when inflammation occurs (Kaltner and Stierstorfer ( 1998) Acta
Anat (Basel) 161: 1-4). Several other animal lectins are expressed in the central and peripheral nervous systems of vertebrates, where the ontogenetic processes, such as axonal growth and fasciculations, neuronal migration, It is thought to be involved in processes involved in cell adhesion and cell recognition mechanisms, such as synapse development and myelination (Zanetta (1998) Acta Anat (Basel) 161: 180-95). Still other animal lectins may be involved in intracellular signaling events at the cell nucleus or at the cell surface by autocrine and paracrine mechanisms (ibid.). Mannose-binding lectin (MBL) is important in stimulating mammalian acute phase responses by mediating the interaction of foreign microbial carbohydrate determinants with serine proteases (ie, MBL-related serine proteases or MASPs). This interaction results in the proteolytic activation of complement component C3 and subsequent phagocytosis or cell lysis (Vasta et al. (1999) Acta
Anat (Basel) 23: 401-20; Zhang et al. (1999) Immunopharmacology 42: 81-
90). In addition, animal lectins have been implicated as mediators of “sugar coding” of cell-cell interactions during embryonic development (Mann and Waterman (199
8) Additional test at 161: 153-61).

[0007] Although lectins may be useful for targeting specific receptors and thus blocking the processes of disease mediated by specific receptors, they have several disadvantages. In particular, for them to have relative specificity, the chemistry of the targeted host cells must be known and the corresponding high affinity receptor binding lectins must be available. Other disadvantages of using lectins to treat or prevent receptor-mediated diseases include their relatively high production costs and difficulty in delivering them to animals.

[0008] Any compound that can bind to a receptor and thus interfere with receptor-mediated interactions would be useful as a drug.

SUMMARY OF THE INVENTION In one aspect, the present invention provides a method of treating a pharmaceutically acceptable carrier comprising an effective amount of a compound of general formula 1
:

[0010]

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(Where X is —OH, —O (aryl), —O (acyl), —O (sulfonyl), —C
Represents N, F, Cl, or Br; Y represents O, S, Se, or NR; Z is an optionally substituted alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, Aralkyl, heteroaralkyl, or — (CH ₂ ) _m —R ₈₀ , where R is each independently hydrogen, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, or — (CH ₂ ) _m —R ₈₀ R ₈₀ each independently represents aryl, cycloalkyl, cycloalkenyl, heterocyclyl, or polycyclyl, and m is an integer in the range from 0 to 8, including 8. And a pharmaceutical composition comprising:

In another aspect, the invention features a method of treating a receptor-mediated disease using a pharmaceutical composition of the invention.

Furthermore, in addition to being effective in treating or preventing a number of receptor-mediated diseases, the compounds of the present invention are derived from the natural zosteric acid obtained from seagrass eel (Zostella marina) Therefore, it should be relatively safe. in addition,
Many of these compounds are small molecules and can be administered orally.

[0014] Other features and advantages of the invention will be apparent from the following detailed description, and from the claims.

DETAILED DESCRIPTION OF THE INVENTION Definitions For convenience, certain terms employed in the specification, examples, and appended claims are described below.

The term “acylamino” is known in the art and has the general formula:

[0017]

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Wherein R ₉ is as defined above, and R ′ ₁₁ is one hydrogen, one alkyl, one alkenyl, or — (CH ₂ ) _m —R ₈ (wherein Medium, m
And R ₈ are as defined above)).

The terms “alkenyl” and “alkynyl” are similar in length, and may be substituted with alkyl as described above, but may include unsaturated aliphatic groups such as those containing double or triple bonds, respectively. Say the group.

The terms “alkoxyl” or “alkoxy” as used herein refer to an alkyl group, as defined above, having an attached oxygen radical. Representative alkoxyl groups include methoxy, ethoxy, propyloxy, t-butoxy, and the like. An "ether" is two hydrocarbons covalently linked by a single oxygen. Accordingly, the substituent of an alkyl that renders that alkyl an ether, for example, -O- alkyl, -O- alkenyl, -O- _{alkynyl, -O- (CH 2) m -R} 8 ( where Shikichu, m and R ₈ is as described above), or is similar to an alkoxyl.

The term “alkyl” is used to refer to saturated alkyl groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. Say the radical. In a preferred embodiment, straight or branched chain alkyl has up to 30 carbon atoms in its main chain (eg, C ₁ -C ₃₀ for straight chain, C ₃ -C _{30 for} branched chain), and more. Preferably 20
Or less carbon atoms. Likewise, preferred cycloalkyls are those having from 3 to 10 carbon atoms in the ring structure, and more preferably having 5, 6 or 7 carbons in the ring structure.

Further, as used throughout the specification and claims, the term “alkyl” (or “lower alkyl”) encompasses both “unsubstituted alkyl” and “substituted alkyl”. As intended, this latter refers to an alkyl moiety having a substituent that replaces a hydrogen on one or more carbons of a hydrocarbon backbone. Such substituents include, for example, halogen, hydroxyl, carbonyl (eg, carboxyl, ester, formyl, or ketone), thiocarbonyl (eg, thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphonate, phosphinate, amino, An amide, amidine, imine, cyano, nitro, azide, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamide, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moiety can be included. Those skilled in the art will appreciate that the moiety substituted on the hydrocarbon chain may itself be substituted where appropriate. For example, substituted alkyl substituents include substituted and unsubstituted forms of amino, azido, imino,
Amide group, phosphoryl group (including phosphonate and phosphinate), sulfonyl group (including sulfate, sulfonamide, sulfamoyl and sulfonate), and silyl group, ether, alkylthio, carbonyl (including ketone, aldehyde, carboxylate, and ester) ), _- CF 3, -CN, may be included so. Representative substituted alkyls are described below. Cycloalkyl is further alkyl, alkenyl, alkoxy, alkylthio, amino alkyl, carbonyl-substituted _alkyl, -CF 3, -CN, optionally substituted with like.

The term “alkylthio” has a sulfur radical attached to it,
Refers to an alkyl group as defined in In a preferred embodiment, the "alkylthio" moiety
Is -S-alkyl, -S-alkenyl, -S-alkynyl, and -S- (CH ₂ )_m-R₈(Where m and R₈Is as defined above)
Represented by one. Representative alkylthio groups include methylthio, ethylthio, and the like.
Is included.

The term “amide” is recognized in the art as an amino-substituted carbonyl,
General formula:

[0025]

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Wherein R ₉ and R ₁₀ are as defined above. Preferred examples of amides will not include imides that may be unstable.

The terms “amine” and “amino” are art-recognized and refer to both unsubstituted and substituted amines, for example, of the general formula:

[0028]

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[0029] (where _Shikichu, R _{9, R 10} and R _'10 are each independently one of hydrogen, an alkyl, an alkenyl, - _(or represents CH ₂₎ m -R _8, or _{R 9} and R _{1 0,} when regarded along with they are attached N atom, which complete a heterocycle having from 8 atoms of four in the ring structure, R ₈ is one of aryl, one of cycloalkyl , One cycloalkenyl, one heterocycle, or one polycycle, and m is zero or an integer from 1 to 8). In a preferred embodiment, only one of R ₉ or R ₁₀ may be a carbonyl, eg, R ₉ , R ₁₀ and this nitrogen do not form an imide. . In an even more preferred embodiment, (depending on and select _{R '10)} R ₉ and _{R 1 0} each individually one of hydrogen, an alkyl, an alkenyl, or - _{(CH 2)} m -R ₈ Represents Thus, as used herein, the term "alkylamine" refers to an amine group, as defined above, having a substituted or unsubstituted alkyl attached thereto, i.e., R _At least one of ₉ and R ₁₀ is one alkyl group.

“Neutral solvent” means at atmospheric pressure, preferably about 25 ° C. to about 190 ° C., more preferably about 80 ° C. to about 160 ° C., most preferably about 80 ° C. to 150 ° C.
A non-nucleophilic reagent having a boiling point above ambient temperature. Examples of such solvents are acetonitrile, toluene, DMF, diglyme, THF or D
MSO.

As used herein, the term “aryl” includes 5-, 6-, and 7-membered single-ring aromatic radicals, which contain from zero to four heteroatoms. Such as benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like. Aryl groups having a heteroatom in the ring structure are sometimes further referred to as "arylheterocycles" or "heteroaromatics". An aromatic ring can be present at one or more ring positions, e.g., halogen, azide,
Alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amide, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclyl, An aromatic or heteroaromatic moiety, —CF ₃ , —CN, and the like, may be substituted with the substituents described above. The term “aryl” further includes two or more rings in which two or more carbons are common to two adjacent rings (these rings are “fused”). And polycyclic systems wherein at least one of the rings is aromatic, for example, the other rings are cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, and / or heterocyclyl.

The term “arylalkyl” as used herein refers to an alkyl group substituted with one aryl group (eg, an aromatic or heteroaromatic group).

As used herein for the purposes of interchangeability, “biological activity” or biological activity ”or“ activity ”or“ biological function ”refers to a compound of the present invention or a fragment thereof. An effector or antigenic function that acts directly or indirectly.

“Bioavailable” is intended to refer to the location or orientation of a compound that is suitable for exerting the biological activity of the compound.

“Biofilm” refers to the accumulation of organisms on one surface. Adult biofilms can include colonies of microorganisms resident on one surface, surrounded by exopolysaccharide.

“Biofilm resistance” or “fouling resistance” refers to the inhibition or reduction of the amount of fouling organisms that attach and / or grow.

As used herein, the term “carbocycle” refers to an aromatic or non-aromatic ring in which each atom of the ring is carbon.

The term “carbonyl” is recognized in the art and has the general formula:

[0039]

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(Where X represents one bond or one oxygen or one sulfur,
And R₁₁Is one hydrogen, one alkyl, one alkenyl,-(CH₂) _m -R₈Or a pharmaceutically acceptable salt;₁₁Is one hydrogen, one atom
Alkyl, one alkenyl or-(CH₂)_m-R₈(Where m and R are ₈ Is the same as defined above)).
is there. X is an oxygen and R₁₁Or R '₁₁If is not hydrogen,
Will represent one "ester". X is one oxygen, R₁₁But
This moiety is referred to herein as a carboxyl group, as defined above.
And especially R₁₁Where is a single hydrogen, this formula represents a "carboxylic acid"
It is. X is an oxygen and R '₁₁If is hydrogen, the formula is
Formate ”. Generally, the oxygen atom in the above formula is replaced by sulfur
In this case, the formula will represent a "thiolcarbonyl" group. X is one sulfur
Yes, R₁₁Or R '₁₁If is not hydrogen, this formula converts the “thiol ester”
It represents. X is one sulfur, R₁₁If is hydrogen, this equation is
All carboxylic acid ". X is one sulfur, R '₁₁Is hydrogen
If so, this formula represents "thiol formate". On the other hand, one X
And R is₁₁If is not hydrogen, the above formula replaces one "ketone" group.
It represents. X is a bond and R₁₁If is hydrogen
The formula represents an "aldehyde" group.

The term “electron-withdrawing group” is art-recognized and refers to the property that one substituent attracts a valence electron from an adjacent atom, that is, the substituent is an adjacent atom. Is electrically negative for The level of the electron withdrawing force can be determined by Hammett's sigma () constant. This well-known constant has been published in
J. March, Advanced Organic Chemistry McGraw Hill Book Company, New York,
(1977 edition) pages 251 to 259. The value of the Hammett's constant is generally negative for an electron donating group ([P] =-0.66 for NH ₂ ), assuming that [P] indicates para-substitution, and Positive in the case of an electron withdrawing group ([P] = 0.78 in the case of a nitro group). Representative electron withdrawing groups include nitro, ketone, aldehyde, sulfonyl, trifluoromethyl, -CN,
Chloride, and so on. Representative electron donating groups include amino, methoxy, and the like.

The term “half-life” refers to the amount of time required for half of a compound to be lost or degraded in the natural process.

As used herein, the term “heteroatom” means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, sulfur and phosphorus.

The term “heterocyclyl” or “heterocyclic group” refers to a 3- to 10-membered ring structure, more preferably a 3-membered ring structure, such that the ring structure contains 1 to 4 heteroatoms. Say 7-member ring. Heterocycles may also be polycyclic. Heterocyclyl groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxatin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole , Indazole, pudding,
Quinolidine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, tinoline, pteridine, carbazole, carboline, phenanthridine, acridine, perimidine, phenanthroline, phenazine, phenalzazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiolane, thiolane, thiolane, thiolane , Piperidine, piperazine, morpholine, lactone,
Includes lactams such as azetidinone and pyrrolidinone, sultam, sultone, and the like. The heterocyclic ring is one or more of one or more substituents as described above,
For example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amide, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, one It may be substituted with heterocyclyl, one aromatic or heteroaromatic moiety, —CF ₃ , —CN, or the like.

Unless the number of carbons is specifically stated otherwise, “lower alkyl” as used herein refers to one to ten carbons, more preferably one to six carbon atoms in its backbone structure. Refers to an alkyl group, as defined above, having Similarly, "lower alkenyl" and "lower alkynyl" have similar chain lengths. Preferred alkyl groups are lower alkyl. In a preferred embodiment, the substituents designated herein as alkyl are lower alkyl.

As used herein, the terms “infectious microorganism” or “infectious agent” cause or contribute to disease (eg, Staphylococcus species (eg, Staphylococcus aureus, Staphylococcus— Bacteria (including Gram-negative and Gram-positive organisms), such as Enterococcus sp. (E. faecalis), Pseudomonas sp. (P. aeruginosa), Escherichia sp. (E. coli), Proteus sp. (P. mirabilis), (Candida) Fungi (including Albicans), (H
Viruses, including IV, HCV, CMV, HBV, and (eg, spirochetes sp)
p, protozoa (such as treponema spp).

As used herein, the term “nitro” refers to —NO ₂ , “halogen” refers to —F, —Cl, —Br, or —I, and the term “sulfhydryl” refers to —SH. and, the term "hydroxyl" means -OH, and the term "sulfonyl" is -SO 2 _- means.

The terms ortho, meta and para are used for 1,2-, 1,3- and 1,4-disubstituted benzenes, respectively. For example, 1,2-dimethylbenzene and ortho-dimethylbenzene are synonymous.

“Pharmaceutically effective amount” refers to an amount suitable to provide a therapeutic effect. Compound poison
Sex and therapeutic efficacy can be determined, for example, by using Ld₅₀(Lethal dose for 50% of the population) and Ed ₅₀ Cells, eg, to determine (therapeutically effective amount for 50% of the population)
Determined by performing standard pharmaceutical procedures on cultured or experimental animals
. The dose ratio between toxic and therapeutic effects is the therapeutic index and the LD₅₀/ ED_{5 0} Can be represented by Compounds that exhibit large therapeutic indices are preferred. Effective dose is
It may vary within a certain range depending on the dosage form used and the route of administration utilized. for
Amount suppresses IC50 (ie half to maximal symptoms) when examined in cell culture
Animal model to achieve a circulating plasma concentration range that includes
It may be made with a tool.

“Pharmaceutically effective carrier” refers to a physiologically acceptable carrier or excipient.
Thus, the compounds and their physiologically acceptable salts and solvates may be administered, for example, by injection, inhalation or insufflation (either through the mouth or nose), orally, buccally, parenterally or rectally. May be formulated for administration by a. For treatment, the compounds of the invention may be formulated for various modes of administration, including systemic and local or local administration. The technology and formulation is generally based on Remington's Pharmaceuticals, Inc. of Mead Publishing, Easton, PA.
Will be found in Sciences. For systemic administration, injection is preferred, including intramuscular, intravenous, intraperitoneal, and subcutaneous. For injection, the compounds of the invention may be formulated as liquid solutions, preferably in physiologically compatible buffers such as Hanks' solution or Ringer's solution. In addition, the compounds may be formulated in solid form and redissolved or suspended immediately before use. Lyophilized forms are also included. For oral administration, the pharmaceutical composition may include, for example, pharmaceutically acceptable excipients, such as a binder (eg, pregelatinized corn starch, polyvinylpyrrolidone or hydroxypropylmethylcellulose), a filler (eg, lactose, Together with a microcrystalline cellulose or calcium hydrogen phosphate, a lubricant such as magnesium stearate, talc or silica, a disintegrant such as potato starch, sodium starch glycolate, or a wetting agent such as sodium lauryl sulfate; It may be in the form of tablets or capsules prepared by conventional means. Tablets may be coated by methods known in the art. Liquid preparations for oral administration may take the form of, for example, solutions, syrups or suspensions, or they may be
It may be provided as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations include, for example, suspending agents (eg, sorbitol syrup, cellulose derivatives or hydrogenated edible oils), emulsifiers (eg, lecithin or gum acacia), non-aqueous vehicles (eg, thiond oil, oily esters). ,
It may be prepared by conventional means together with pharmaceutically acceptable additives such as ethyl alcohol or fractionated vegetable oils) and preservatives (for example methyl or propyl p-hydroxybenzoate or sorbic acid). . Formulations further include buffer salts, flavors,
Coloring and sweetening agents may be included as appropriate. Preparations for oral administration may be suitably formulated to give controlled release of the active compound. For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner. For administration by inhalation, the compounds employed in accordance with the present invention are pressurized with a suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. It is also convenient to deliver them in the form of an aerosol spray from a pack or nebulizer. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin for use in an inhaler or insufflator may be formulated containing a powder of a mixture of the compound and a suitable powder base such as lactose or starch. The compounds may be formulated for parenteral administration by injection, eg, by bolus injection or continuous infusion. Preparations for injection
It may be presented in unit dosage form with added additives, for example, in ampoules or in multi-dose containers. The compositions may be in the form of suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and / or dispersing agents. May be included. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, such as sterile, non-pyrogenic water, before use. Additionally, the compounds may be formulated in rectal compositions such as suppositories or fixed enemas, eg, containing conventional suppository bases such as cocoa butter or other glycerides.
In addition to the formulations described previously, the compounds may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated with a suitable polymeric or hydrophobic material, such as, for example, as an emulsion in an acceptable fat or oil, or an ion exchange resin, or, for example, with limited solubility. They may be formulated with limited soluble derivatives such as salts. Further, systemic administration may be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are known in the art, and include, for example, for transmucosal administration bile salts and fusidic acid derivatives. In addition, a surfactant may be used to facilitate penetration. Transmucosal administration may be through nasal sprays or using suppositories. For topical administration, the oligomers of the invention may be formulated into ointments, salves, gels, or creams as generally known in the art. A cleaning solution may be used topically to treat the injury or inflammation and speed healing.

“Phosphoryl” generally has the formula:

[0052]

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Wherein Q ₁ represents S or O and R ₄₆ represents hydrogen, one lower alkyl or one aryl. For example, when substituting an alkyl or the like, the phosphoryl group of the phosphorylalkyl has the general formula:

[0054]

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Wherein Q ₁ represents S or O, and each R ₄₆ independently represents hydrogen, one lower alkyl or one aryl, and Q ₂ represents O, S or N. Can be represented. When Q ₁ is an S, the phosphoryl moiety is “phosphorothioate”.

“Polar neutral solvents” include, for example, DMF, acetonitrile, diglyme, DM
Means a polar solvent as defined above, such as SO or THF, which has no hydrogen exchangeable compound of the invention during the reaction.

“Polar solvents” include, for example, DMF, THF, ethylene glycol dimethyl ether (DME), DMSO, acetone, acetonitrile, methanol, ethanol, isopropanol, n-propanol, t-butanol, 2-methoxyethyl ether, and the like. It means a solvent having a dielectric constant (ε) of 0.9 or more. Preferred solvents are DMF, DME, NMP, and acetonitrile.

The term “polycyclyl” or “polycyclic group” is used when two or more carbons are common to two adjacent rings, such as when two or more rings are “fused rings”. Refers to two or more rings (eg, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl and / or heterocyclyl, etc.). Rings that are joined through non-adjacent atoms are termed "bridged" rings. Each of the polycyclic rings is, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amide, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, Ketones, aldehydes, esters, one heterocyclyl,
An aromatic or heteroaromatic moiety of, -CF 3, _-CN, may be substituted with such substituents as described above, such like.

The term “protecting group” as used herein refers to a temporary modification of one potentially reactive functional group, such as to protect it from unwanted chemical transformations. Examples of such protecting groups include esters of carboxylic acids, silyl ethers of alcohols,
And there are acetals and ketals of aldehydes and ketones, respectively. This field of protecting group chemistry has been replicated (Greene, TW; Wuts, PGM Protective Gr.
oups in Organic Synthesis, 2nd ed .; Wiley: New York, 1991).

As used herein, “receptor-mediated disease or condition” refers to any of a wide variety of diseases or conditions that involve receptor-mediated pathways. Particularly preferred diseases or conditions are those involving the sulfate receptor-mediated pathway. Examples include infections (by viruses, bacteria, fungi, protists, prions), cancer, fertilization and blood clotting related diseases.

As used herein, the term “substituted” is considered to include all acceptable substituents on organic compounds. In a broad sense, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic groups of organic compounds. Examples of substituents include, for example, those described above. The permissible substituents can be one or more and the same or different for suitable organic compounds. For purposes of the present invention, a heteroatom such as nitrogen may have a hydrogen substituent and / or any acceptable substituent of an organic compound described herein that satisfies the valence of the heteroatom. Is also good. The present invention is not intended to be limited in any manner by the permissible substituents of organic compounds.

By “substituted” or “substituted with” is meant that such substitutions are in accordance with the possible valencies of the substituted atoms and substituents, resulting in, for example, rearrangements, It will be understood that the implicit assumption is made that stable compounds can be obtained, such as compounds that do not spontaneously perform transformations such as cyclization, removal, and the like.

The term “sulfate” is recognized in the art and has the general formula:

[0064]

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Where R ₄₁ is as defined above.

“Sulfate binding moiety” refers to a moiety that can bind or associate with a sulfate group or a sulfonate group.

The term “sulfonate” is recognized in the art and has the general formula:

[0068]

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Wherein R ₄₁ includes a moiety that can be represented by one electron pair, hydrogen, alkyl, cycloalkyl, or aryl.

As used herein, the terms “sulfoxide” or “sulfinyl”
General formula:

[0071]

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(Wherein, R ₄₄ represents a moiety which can be represented by any one of hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aralkyl, and aryl).

The alkenyl group and the alkynyl group may be similarly substituted to give, for example, aminoalkenyl, aminoalkynyl, amidoalkenyl, amidoalkynyl, iminoalkenyl, iminoalkynyl, thioalkenyl, thioalkynyl, carbonyl-substituted alkenyl or alkynyl. Can be generated.

“Sustained release” or “controlled release” refers to a relatively constant or prolonged release of a compound of the present invention from one surface, with a central location around the compound of the present invention. This can be achieved by using a reservoir system in which is surrounded by a porous membrane or layer, or a diffusion system including a matrix system in which the present compound is distributed throughout an inert matrix. Materials that may be used to form the matrix include:
Examples include silicones, acrylates, methacrylates, vinyl compounds such as polyvinyl chloride, olefins such as polyethylene or polypropylene, fluoropolymers such as polytetrafluoroethylene, and polyesters such as terephthalate. The diffusion system may be formed into a film or other layered material and then placed in adhesive contact with a structure intended for underwater use. Optionally, the compound of the present invention may be mixed with a resin such as polyvinyl chloride, and then molded into a molded product.The molded product thus incorporates the compound into the compound, or the compound or the compound. It forms a structure with a porous matrix that allows the functional part to diffuse into the surrounding environment. Furthermore, if micro-encapsulation technology is used,
Sustained local release of the compounds of the present invention can also be maintained. Furthermore, the microencapsulation method may be used to improve stability. The encapsulated product may, for example, take the form of a sphere, an aggregate of core material embedded in a continuum of wall material, or a capillary. The core material of the microcapsules containing the sulfated AF agent may be in the form of droplets, emulsions, suspensions of solids, solid particles, or crystals. One skilled in the art will recognize that many materials suitable for use as coating materials for microcapsules, including but not limited to organic polymers, hydrocolloids, lipids, fats, carbohydrates, waxes, metals, and inorganic oxides You will know.
Silicone polymers are the most suitable microcapsule coating materials for treating surfaces. Microencapsulation techniques are known in the art and are incorporated herein by reference. Encyclopedia of Polymer Science and Engineering, Vol. It is described after pages 9, 724.

As used herein, the term “treating” is intended to include curing and ameliorating at least one symptom of the condition or disease.

The abbreviations Me, Et, Ph, Tf, Nf, Ts, Ms and dba stand for methyl, ethyl, phenyl, trifluoromethanesulfonyl, nonafluorobutanesulfonyl, p-toluenesulfonyl, methanesulfonyl, and dibenzylideneacetone, respectively. Represents A more comprehensive list of abbreviations used by organic chemists in the art is
As found in the first edition of each volume of the Journal of Organic Chemistry, this list is typically represented by a table entitled Standard List of Abbreviations. The abbreviations included in the above list, and all abbreviations used by organic chemists in the art, are hereby incorporated by reference.

For the purposes of the present invention, chemical elements are defined in the Handbook of Chemistry.
And Physics, 67th edition, 1986-87, based on the Periodic Table of Elements in the CAS version inside the cover. Further, for the purposes of the present invention, the term "hydrocarbon" is considered to include any acceptable compound having at least one hydrogen and one carbon atom. In a broad aspect, the acceptable hydrocarbon includes acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic, substituted or substituted. Organic compounds that may not be present are included.

Pharmaceutical Compositions of the Invention In some embodiments, a pharmaceutical composition of the invention comprises a pharmaceutical carrier and a compound of general formula 1:

[0079]

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(Where X is —OH, —O (aryl), —O (acyl), —O (sulfonyl), —C
Represents N, F, Cl, or Br; Y represents O, S, Se, or NR; Z is an optionally substituted alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, Aralkyl, heteroaralkyl, or — (CH ₂ ) _m —R ₈₀ , where R is each independently hydrogen, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, or — (CH ₂ ) _m —R ₈₀ R ₈₀ each independently represents aryl, cycloalkyl, cycloalkenyl, heterocyclyl, or polycyclyl, and m is an integer in the range from 0 to 8, including 8.) An effective amount of the compound. Other suitable compounds are salts of the compounds of general formula 1.

Particularly stable compounds are those represented by general formula 1 and the attendant definitions, wherein X is —OH, F,
Cl (or Br)).

In another preferred embodiment, the compositions of the present invention comprise a compound represented by general formula 1 and the attendant definitions, wherein Y represents O.

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions wherein the formula
Wherein Z is an optionally substituted alkyl, aryl, or-(CH₂)_m-R ₈₀ Represents a compound).

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein Z is an optionally substituted alkylphenyl, heteroalkylphenyl,
Arylphenyl or heteroarylphenyl).

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein Z is 4- (2-methylpropyl) phenyl, 4- (1,1-dimethylethyl). ) Phenyl,
4- (1,1-dimethylpropyl) phenyl, 4-pentylphenyl, 4- (1-methyl-1-phenylethyl) phenyl, or 4- (1-methylheptyl) phenyl) Including.

In some embodiments, the compositions of the present invention are compounds represented by general formula 1 and the attendant definitions, wherein R represents H or alkyl.

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH, F, Cl, or Br, and Y represents O. And a compound represented by the formula:

In some embodiments, the compositions of the present invention include compounds of general formula 1 and the attendant definitions, wherein X represents —OH or Cl, and Y represents O. including.

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH, F, Cl, or Br; and Z is optional. containing the compound represented by represents a _{(CH 2) m -R 80)} - substituted alkyl, aryl, or.

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH or Cl, and Z is an optionally substituted alkyl,
Containing _(CH 2) a compound represented by m _-R represents a _{80) -} aryl, or.

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH, F, Cl, or Br; and Z is optional. Represents a substituted alkylphenyl, heteroalkylphenyl, arylphenyl, or heteroarylphenyl).

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH or Cl, and Z is an optionally substituted alkylphenyl. , Represents a heteroalkylphenyl, an arylphenyl, or a heteroarylphenyl).

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH, F, Cl, or Br, and Z is 4- (2 -Methylpropyl) phenyl, 4- (1,1-dimethylethyl) phenyl, 4- (1,1-dimethylpropyl) phenyl, 4-pentylphenyl, 4- (1-methyl-1-phenylethyl) phenyl, or
4- (1-methylheptyl) phenyl).

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH or Cl, and Z is 4- (2-methylpropyl) phenyl ,Four
-(1,1-dimethylethyl) phenyl, 4- (1,1-dimethylpropyl) phenyl, 4-pentylphenyl, 4- (1-methyl-1-phenylethyl) phenyl, or 4- (1-methylheptyl) ) Represents phenyl).

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein Y represents O and Z is optionally substituted alkyl, aryl, or — (CH ₂ ) _m —R ₈₀ ).

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein Y represents O, and Z is optionally substituted alkylphenyl, heteroalkyl. Represents phenyl, arylphenyl, or heteroarylphenyl)
And a compound represented by the formula:

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein Y represents O and Z is 4- (2-methylpropyl) phenyl, 4- (1,1-dimethylethyl) phenyl, 4- (1,1-dimethylpropyl) phenyl, 4-pentylphenyl, 4- (1-methyl-1-phenylethyl) phenyl, or 4- (1-methylheptyl) Phenyl).

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH, F, Cl, or Br, Y represents O, and Z is alkyl substituted in accordance with the selection, aryl, or _- includes a compound represented by represents a _{(CH 2) m -R 80)} .

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH or Cl, Y represents O, and Z is optional. containing the compound represented by represents a _{(CH 2) m -R 80)} - alkyl substituted Te, aryl, or.

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH, F, Cl, or Br, Y represents O, and Z represents optionally substituted alkylphenyl, heteroalkylphenyl, arylphenyl, or heteroarylphenyl).

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH or Cl, Y represents O, and Z is optional. Represents a substituted alkylphenyl, heteroalkylphenyl, arylphenyl or heteroarylphenyl).

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH, F, Cl, or Br, Y represents O, and Z is 4- (2-
Methylpropyl) phenyl, 4- (1,1-dimethylethyl) phenyl, 4- (1,1-dimethylpropyl) phenyl, 4-pentylphenyl, 4- (1-methyl-1-phenylethyl)
Phenyl or 4- (1-methylheptyl) phenyl).

In some embodiments, the compositions of the present invention have general formula 1 and the attendant definitions, wherein X represents —OH or Cl, Y represents O, and Z represents 4- ( 2-methylpropyl) phenyl, 4- (1,1-dimethylethyl) phenyl, 4- (1,1-dimethylpropyl)
Phenyl, 4-pentylphenyl, 4- (1-methyl-1-phenylethyl) phenyl, or 4- (1-methylheptyl) phenyl).

Those skilled in the art will appreciate that the compositions of the present invention can maintain AF activity if necessary to optimize the overall chemical properties of that particular compound for a particular application. It will be appreciated that can be changed with. For example, the length of the alkyl chain can be extended or shortened to control the rate of dissolution of the compound from a structure or coating. Alternatively, additional functional groups are added to the alkyl chain,
The chemistry of the molecule can be further modified.

Biological Activity of Compounds of the Invention Compounds of the invention have been shown to provide one or more plant and animal lectin-like activities. Lectins bind to cell surface proteoglycans, which function in the attachment of pathogens such as viruses and bacteria. Thus, the lectin-like activity of the compounds of the present invention is useful in treating and preventing infections and other receptor-mediated diseases and conditions.

The ability of the compounds to bind to specific cell surface sites is important in activating or antagonizing some cell surface interactions affected by animal or plant lectin proteins. This extracellular polysaccharide produced by fouling organisms is often highly sulfated, and these sulfates play an important role in polymerization (eg, glue / gel formation) (Wetherbee et al., (1998 ) J. Phycol.
34: 9-15; Vreeland et al., (1998) J. Phycol. 34; 1-8).

As shown in the examples below, the compounds of the present invention inhibit these glue / gel interactions. Thus, the compounds of the present invention have many practical uses. Based on the wide range of activities exhibited, the compounds of the invention do not require the precise nature of host receptors involved in the disease process to be determined. Furthermore, the sulfated compounds of the present invention provide a general class of suitable lectin-like therapeutics that do not require the identification of individual receptor-specific lectin species.

In addition, lectins are involved in a wide range of processes, and thus the lectin-like activity of the zosteric acids and related sulfated compounds of the present invention provides a corresponding broad use of the present invention. For example, the involvement of animal lectins in the activation of host complement suggests that zosteric acid-related compounds according to the present invention can be used to treat and prevent undesirable inflammatory diseases and conditions. In addition, the zosteric acids and related compounds of the present invention have some unique properties that are useful in certain applications, including anticoagulant activity similar to that of heparin.

Another important property of the compounds of the present invention is their ability to affect the aggregation of bacterial and mammalian cells. The effect of the compound on cell aggregation involves blocking specific cell surface receptors and blocking other activations, such as those involved in extracellular surface attachment and mediating fouling. May. Thus, the compounds of the present invention have numerous activities of naturally-occurring proteins and glycoproteins that bind to sites on the surface of a single cell and affect cell-to-cell interactions. . Cellular hemagglutinin-like aggregation activity of the compound of the present invention, for example,
It may be useful to promote wound healing.

As shown in the examples below, the compounds of the present invention have been found to have several anticoagulant activities, such as those possessed by the sulfated mucopolysaccharide heparin.
Heparin is thought to inhibit the coagulation cascade by binding to and activating antithrombin III, which in turn forms an irreversible complex with thrombin. It is a plasma protein that inactivates thrombin. This complex is similar to the acyl-enzyme complex formed between trypsin and pancreatic trypsin inhibitor. Heparin is produced by mast cells near the vessel wall and acts as an anticoagulant by increasing the rate of irreversible complex formation between thrombin and antithrombin III. Antithrombin III may also be used, for example, for factors IXa, Xa, and XIa
Inhibits other proteolytic components of the coagulation cascade.

Despite the low molecular weight activity, the zosteric acid molecule is much smaller than that observed for heparin. Heparin is effective in preventing clot formation even at concentrations much greater than 0.1 mg / ml, while zosteric acid is 10 mg / ml.
Only effective at concentrations above / ml. The effect of heparin-like anticoagulants is strongly linked to size, with higher molecular weight molecules being more effective. Therefore, among the compounds of the present invention, those having a higher molecular weight must be more effective.

Heparin has been widely used in the prevention and treatment of deep vein thrombosis. However, heparin has several limitations. For example, heparin, a relatively large molecule, has been shown to have limited efficacy in inhibiting thrombin activity incorporated into fibrin clots. In addition, heparin has a short intravenous half-life. Thus, in some applications, the use of the compounds of the present invention may be advantageous in providing certain anticoagulant prophylaxis or treatment. Further, the compounds of the present invention can be formulated as covalent conjugates derived for the treatment and prevention of coagulation conditions. For example,
Covalent forms of the antithrombin-heparin conjugate have proven to be more effective at reducing coagulation weight in vivo than therapies combining thrombin and heparin. Accordingly, the present invention provides methods and reagents for producing an inducing component of the fibrin coagulation cascade, including antithrombin.

While the compounds of the invention may be delivered orally, heparin must be delivered intravenously. Furthermore, the compounds of the present invention have heparin-like properties and are particularly suitable for incorporation into medical materials where anticoagulant action is desired.

As shown in the examples below, the compounds of the present invention have been found to further inhibit fertilization, so the compounds of the present invention can be used as contraceptives. Birth control pills, including those recited in the claims, are typically formulated as coatings, foams, jellies and suppositories which are reapplied as described above, or, optionally, a prophylactic product such as a condom or May be formulated as a semi-permanent coating used on the diaphragm). Alternatively, during the manufacture of such a product, the product may be compounded with a sulfate ester.

The following examples will further illustrate the invention, but these examples should not be taken as limiting. It is expressly hereby stated that the contents of all cited references, including documents cited throughout this patent application, issued patents and published patent applications, have been incorporated with their references.

EXAMPLES Example 1 Inhibition of Surface Adhesion of Marine Bacteria by Alkyl Sulfate Octyl Sulfate
Alkyl sulphate surfactants with a wide range of industrial uses and are manufactured by several large chemical companies. To investigate the potential AF properties of the octyl sulfate octyl sulfate, it was incorporated into an inert coating material, which was then coated on one surface and the surface was exposed to conditions that helped form a marine algal biofilm.

Materials and Methods Pure octyl sulfate was recovered by subjecting a 30% (w / v) aqueous solution of octyl sulfate (manufactured by Stepan Chemical Co.) to an air stream at room temperature and evaporating it to dryness.
(Fig. 1). The dried octyl sulfate was added to RTV-11 under a load of 25% (wt / wt).
Incorporated into silicone polymer (RTV silicone, catalyst and primers were obtained from General Electric). The mixture was applied to three glass slides pre-treated with a silicone primer and cured to dryness. Three pre-treated glass slides coated with pure RTV-11 served as control with no agent. After complete drying, the absorbency of each slide was measured using a Shimadzu UV-2101 spectrophotometer fitted with an integrating sphere. The slides were then placed in a tank running fresh seawater and incubated outdoors in natural sunlight for 26 days. The water temperature was nominally 15 ° C. Biofilm accumulation as determined by spectrophotometry was measured periodically for each slide. The relative algal biomass was calculated as the ratio of the absorption at 680 nm affected by chlorophyll α to that at 750 nm, a wavelength not absorbed by chlorophyll, to correct for differences in turbidity and scattering of the separate slides.

Results As shown in FIG. 1, octyl sulfate, incorporated into RTV-11 silicone and then coated on glass slides, significantly inhibited the formation of natural seaweed biofilms in natural seawater. After incubation in flowing seawater for 26 days, the growth of algal biofilms on the coating containing octyl sulfate is one-fifth that of the control without octyl sulfate, indicating that octyl sulfate has strong AF activity. Was showing.

Experiments were performed to evaluate the ability of the sulfate molecules octyl sulfate and methyl sulfate to inhibit the adhesion of the marine bacteria Oceanosprilam and Alteromonas-Atlantica to glass surfaces.

Materials and Methods Oceanosprilam Adhesion Test Each test consisted of a control set (no sulfate) and a sample set containing test molecules. The first test group is a control sample set,
It consisted of a zosteric acid (5 mM) sample set and an octyl sulfate (5 mM) sample set. The second test group consisted of a control sample set, a zosteric acid (5 mM) sample set, and a methyl sulfate (5 mM) sample set. The sample set consisted of five 50 mL sterile centrifuge tubes containing 50 mL, each containing a glass microscope slide and 1 × 10 ⁶ cells of the Oceanosprilam culture strain in which sulfate was dissolved. / ML of artificial seawater (ASTM-American Society for Testing and Materials) inoculated to a volume of / mL. The sample set was incubated at 23 ° C. with shaking so that the slide surface was horizontal. Over a 6 hour period, individual test tubes were removed from the sample set and examined for microbial attachment.

Alteromonas-Atlantica Adhesion Test This test consisted of a control sample set, a zosteric acid (5 mM) sample set, an octyl sulfate (5 mM) sample set, and a methyl sulfate (5 mM) set. The sample set consisted of six 60 mL sterile centrifuge tubes. Each tube was inoculated with a glass microscope slide and an Alteromonas-Atlantica culture to an initial cell density of 1 × 10 ⁶ cells / mL, and 50 mL of modified agent dissolved in 50 mL. ASTM Seawater (American Society for Testing and Materials (1986)
D1141-86, ASTM, Philadelphia, PA). Although this modified seawater was composed of normal ASTM seawater components, the amount was such that the carbon was sufficient for deposition but did not achieve significant cell growth.
Glycerol, a carbon source, is typically only one thousandth of its strength,
0.1 L / L instead of L / L and no amino acid source (casamino acid).

Determination of Bacterial Attachment The sample was removed from the shaker and 1 mL of 50 × acridine orange dye (0.5 g / L aqueous solution of acridine powder) was added to the test tube. The dye was allowed to react for 4 minutes. The slides were then taken, fitted with a long cover slip and immediately counted on an epifluorescent microscope equipped with a 100 × (oil) objective on the underside of the slide. The size of the counting visual field was 10 × 10 μm. A total of 20 counts were performed per slide and averaged to give the number of cells per square μm, which was converted to the number of cells per square mm. 10% error
This is the standard accepted error when directly counting bacterial cells.

Results As shown in FIG. 2, when octyl sulfate or methyl sulfate is present in the medium,
Bacteria attached to glass slides were significantly reduced compared to controls without any sulfate molecules. Methyl sulphate inhibits the adhesion of oceanosprilam to a similar extent as the proven AF agent zosteric acid, and each compound reduces bacterial adhesion by approximately one-half compared to controls. Reduced. As shown in FIG. 3, the extent to which octyl sulfate inhibited the attachment of Oceanosprilam was even greater than that of zosteric acid.

As shown in FIG. 4, in the presence of dissolved zosteric acid, octyl sulfate, or methyl sulfate, there was a significant decrease in adhesion of the marine bacterium Alteromonas-Atlantica compared to controls. The presence of methyl sulphate had the most dramatic effect on the attachment, but the attachment was constant at 150,000 cells / mm ² after 120 minutes, while the control had 700,000 cells / mm ^2. It was over mm ^2. Octyl sulfate also inhibited adhesion, suggesting a slightly higher inhibitory activity than zosteric acid.

Example 2: Inhibition of Fungal Surface Attachment and Hyphal Development [0125] To determine the effect of sulfate on fungal biofouling, zosteric in inhibiting fungal aureobasidium-pullance from adhering to surfaces. The ability of the acid was examined.

Materials and Methods Aureobasidium-pullans (ATCC 34261) was grown on potato starch agar and subjected to the ASTM G21-90 protocol (American Society for Testing and Materials (1986) D1141-8).
6, ASTM, Philadelphia, PA). The resulting spore suspension was used to inoculate a liquid culture tube containing 35 mL of growth medium (nutrients supplemented with 5 mM sucrose) and 15 mM zosteric acid. A medium without zosteric acid was made as a control. Sterile microscope slides were added to each tube, and the tubes were sealed and placed on a rotary shaking table at room temperature. For one test tube, the slides were removed and collected daily by counting the number of spores attached by direct microscopic counting as described above.

Results Fungal spores were observed to grow in both the presence and absence of zosteric acid, as indicated by cloudiness of all test tubes after day one. However, as shown in FIG. 5, the presence of zosteric acid prevented the fungus from adhering to the glass slide. A. After incubation with pullulans for 5 days, less than 20 germinated fungal colonies / mm ² were observed on slides incubated in the presence of added zosteric acid, which was 600 In contrast to that there were more than one germinated fungal colony / mm ² .
Furthermore, the fungal colonies in the culture medium without zosteric acid were composed of multicellular (> 20 cells) filaments, indicating hyphal growth. In contrast, the colonies in the zosteric acid-treated cultures were generally small and round, with no evidence of filament growth or hyphal development.

Example 3: Sulfate Binds on the Cell Surface of Biofouling Organisms To investigate the mechanism behind the AF activity of sulfate, a polyclonal antibody specific to sulfate zosteric acid was generated ( Berkeley, CA, manufactured by Babco). Preliminary testing of these antibodies for cross-reactivity to related compounds without sulphate groups (cinnamic, ferulic, and coumaric acids) showed no cross-reactivity and the specificity recognized by this antibody. It was suggested that the domain would probably contain a sulfate group. Next, using these antibodies, it was examined whether the sulfate ester AF agent zosteric acid directly binds to fouling organisms.

The marine bacterium Schwanella putritifaciens was grown in culture containing zosteric acid and subsequently examined for bound zosteric acid using immunosilver staining with the antibodies described above. S. with immunoprobe. Electron microscopy of the putrifaciens detected zosteric acid molecules bound to the bacterial surface. Furthermore, it was observed that zosteric acid was present at a high incidence even at the cell adhesion site. In contrast to these aggregation sites, the majority of the continuous boundaries between daughter cells in the cell surface and in dividing chains, as evidenced by the absence of immunosilver staining, indicate that bound zosteric acid There was no evidence. These results suggest that sulfate binds to the surface of bacterial cells, including a sulfate binding site,
This indicates a potential relationship with the bacterial aggregation site.

Example 4 Zosteric Acid Promotes Bacterial Aggregation To further investigate the role of sulfate in aggregation, the ability of sulfate to promote bacterial cell aggregation was examined. Log phase cultures grown in the presence of zosteric acid were observed by spectrophotometry (OD ₆₀₀ ) for growth and aggregation in the presence of increasing amounts of zosteric acid.

Materials and Methods Cell Surface Binding Assay The marine bacterium Schwanella-Putrifaciens was
Grow in seawater broth in the presence of 6 mM zosteric acid. The dense log phase cells were harvested after 5 hours of growth and harvested with 0.5 × Karnovski's fixative (2%
Formaldehyde, 2.5% glutaraldehyde, 0.05M sodium cacodylate, 0.25M sucrose, pH 7.4) for 2 hours, and then stored in a cacodylate buffer (0.05M Sodium cacodylate, pH 7.4
). Cells were prepared for electron microscopy using the immunosilver staining technique (Harlow, E. and Laine, D., Antibodies, A Laboratory Manual, Cold Spring
Harbor Laboratory, 359-421; Roth et al., J. Histochem. Cytochem. 26: 10
74-1081 (1978)). The primary antibody used in this study was an anti-zosteric acid polyclonal antibody (Babco, Richmond, CA).

Bacterial Aggregation Assay A log phase culture of Schwanella putritifaciens was
Grow in complete seawater medium containing a range of zosteric acid at concentrations up to 20 mM. At 8 hours, cultures were counted for surviving colony forming units.

Results At zosteric acid concentrations up to 16 mM, S. cerevisiae in liquid cultures was increased. Although there was no inhibition of P. trifaciens growth, the presence of zosteric acid caused S. cerevisiae to grow in a concentration-dependent manner. There was significant aggregation in the putrefaciens. The observed aggregation was also visible to the naked eye and was detected more quantitatively as a decrease in optical density absorbance in the culture solution containing zosteric acid (FIG. 6). As a result of counting viable colony forming units at 8 hours, there was no difference in cell density between the different cultures, and thus the observed difference in absorbance was due to the rate of growth (cell division) between the cultures. The difference was not due to differences in bacterial aggregation. Thus, zosteric acid promoted cell aggregation but did not affect cell growth.

Example 5 Zosteric Acid Binds to Heparin-Sensitive Sites Hemagglutination to determine the ability of sulfate to mediate interactions between biological surfaces involved in erythrocyte aggregation and blood clotting Assays and clot formation assays were performed with sulfated zosteric acid.

Materials and Methods Hemagglutination Assay Washed equine erythrocytes suspended in 1 mg / mL sodium citrate were placed in microtiter plates designed with hemispherical bottoms of wells. Negative controls (no zosteric acid) were diluted in isotonic saline. Cells treated with zosteric acid were treated with 0.005 to 5.0 mg / mL.
Was diluted with saline containing eight concentrations of zosteric acid. Positive controls were exposed to the same range of high molecular weight heparin sulfate concentrations.

Coagulation Assay Regarding prothrombin clotting time, a clotting time assay was performed using a commercially available kit (manufactured by Sigma Chemical Co.). Blood cells were removed by centrifugation from 30 mL of human whole blood obtained by venipuncture, and serum was collected. Zosteric acid and high MW heparin were added to separate aliquots of this serum to bring the concentration to 0.
To 5.0 mg / mL. Clotting times were duplicated for each concentration based on the protocol provided with the kit.

Results In coagulation experiments, equine erythrocytes were significantly aggregated at concentrations as low as 0.175 mg / mL zosteric acid. In contrast, in the presence of high molecular weight heparin, there was visible aggregation only at concentrations above 0.75 mg / mL. The result is
The monomeric pair, zosteric acid, has been shown to be eight times more reactive with cell surface glycoproteins and polysaccharides involved in cell aggregation than high molecular weight heparin.

Zosteric acid is also effective in preventing clot formation as measured by the prothrombin clotting time assay (FIG. 7), but its activity is significantly less than that observed for heparin. Was. Heparin was effective at preventing coagulation at concentrations much lower than 0.1 mg / ml, whereas zosteric acid was only effective at concentrations above 10 mg / ml. Heparin-like anticoagulant effects are strongly related to size, with higher molecular weight molecules being more effective. Thus, it is not surprising that low molecular weight zosteric acids are much less effective than high molecular weight heparin in mediating clot formation. Derivatives of zosteric acid or another sulfate ester of higher molecular weight may be more effective. However, these results indicate that zosteric acid
It suggests interacting with cell surface glycoproteins and / or polysaccharides in a manner similar to heparin.

Example 6 Zosteric Acid Blocks Fertilization The above data suggests that sulfate interacts with a wide range of sulfate-binding receptors, from bacteria to mammalian erythrocytes. Sperm and egg cell fusion in invertebrate and mammalian systems also appear to be mediated by organic sulfate molecules such as the polysaccharides fucose sulfate and heparin. With this in mind, the following experiment was initiated to elucidate the potential AF properties of sulfate during fertilization.

The ability of sulfate to block sperm-egg fusion was detected and quantified using a simple sea urchin assay. Sea urchin sperm were added to freshly harvested eggs in the presence and absence of increasing amounts of sulfated zosteric acid, and the eggs were scored for successful fertilization.

Materials and Methods Fertilization Assay A 0.5 M KCL solution was injected into healthy sea urchins for oviposition. Freshly collected fresh eggs are gently washed and filtered seawater (FSW, p
H was resuspended in 8.29 and aliquoted into separate tubes for fertilization assays. Zosteric acid was added to each test tube taken from the concentrated strain dissolved in FSW (pH 8.2) while further adding FSW so that the volume in each test tube was constant. An equal volume of sperm was added to each tube and percent fertilization was determined by direct microscopic counting. Eggs with raised zygote were scored as fertilized. Assays were performed at a limited concentration of sperm numbers such that 95-99% fertilization occurred in the absence of zosteric acid.

Sea Urchin Egg Aggregation Assay Agglutination of non-fertilized eggs by bindin was evaluated in the range of concentrations of zosteric acid shown in Table 1. Freshly released eggs were suspended in production seawater (pH 5) for 5 minutes to remove the outer jelly membrane, and then washed 5 times with normal FSW (pH 8.2). The eggs were then transferred to plastic petri dishes containing a range of zosteric acid concentrations and incubated for 15 minutes. Purified bingin (Stanford University D. Ieper) was added to the eggs at 1.2
To 12 μg / mL. The mixture was gently agitated on a rotary shaker for 5 minutes and visually examined for aggregation. Bovine serum albumin (BSA
) Was used in another assay to control non-specific aggregation of de-jellyed eggs.

Point Blot Assay Serial dilutions of purified bindin, covalently bound zosteric acid-BSA molecule, and unbound free zosteric acid and BSA
The mixture was dropped on a nitrocellulose membrane with a pipette and air-dried. next,
The reactivity between the blotted substrate and the polyclonal anti-zosteric acid antibody was determined using standard immunoblot techniques. The membrane was blocked with a blot (1% skim milk powder in phosphate buffered saline (PBS)) for 1 hour before probing.
Probing was performed on the blot for 1 hour. The primary antibody was an anti-zosteric acid antibody used at a dilution of 1: 1000. The secondary antibody was an alkaline phosphatase-conjugated goat anti-rabbit (Southern Biotechnology Association), used at a dilution of 1: 1000. Rinsing with PBS-Tween was performed in triplicate between probings. The blot was prepared using a color reaction buffer (100 mM Tris, pH 9.5, 100 mM NaCl, 5 mM MgCl 2, 50 mg).
/ ML of nitro blue tetrazolium (manufactured by Sigma) and 50 ml / mL of 5-bromo-4-chloro-3-indolyl phosphate (BCIP, manufactured by Sigma) for 20 minutes. Then stop the membrane with stop buffer (10 mM Tris, pH 6.0, 5 mM
(EDTA) for 1 hour, then transferred to fresh water, left overnight and then dried.

Results As shown in FIG. 8a, zosteric acid had a dose dependent effect on fertilization of sea urchin eggs. At concentrations higher than 0.5 mg / mL (1.5 mM), fertilization was completely blocked. Unfertilized eggs that received the highest zosteric acid treatment were re-exposed to fresh sperm after being washed with seawater, resulting in fertilization of all eggs. This result demonstrates that inhibition of zosteric acid is reversible. No effect of the presence of zosteric acid on sperm viability or motility was detected.
Spermatozoa exposed to zosteric acid were observed to swim vigorously through the jelly layer around the egg without attaching to the surface of the egg or raising the fertilizing membrane of the egg. Further support the conclusion that the antifouling action of zosteric acid is mediated through the inhibition of sperm-egg adhesion.

The effect of zosteric acid (1 mg / mL) on fertilization inhibition was compared to equal mass concentrations of coumaric acid (unsulfated zosteric acid precursor) and high MW heparin. While the presence of Kumar had no effect on egg fertilization, the presence of heparin reduced fertilization by almost 50%. Zosteric acid is at least twice as effective as heparin in inhibiting fertilization, and at this concentration
It was reduced to 1% (FIG. 8b).

The ability of zosteric acid to compete for binding to the sulfate receptor on the egg surface was examined in an egg aggregation assay. These experiments examined the ability of zosteric acid to buffer the Bindin molecule from binding to unfertilized sea urchin eggs. When binzin is added to unfertilized eggs, these eggs aggregate by cross-linking on sulfate receptors on the egg surface. Addition of zosteric acid inhibited this aggregation in a dose-dependent manner (Table 1), suggesting that there may be a competitive interaction between bindin and zosteric acid over the sulfate receptor site on the egg surface. It was suggested.

[0147]

[Table 1]

Antibodies specific for zosteric acid (described above) showed strong cross-reactivity with the Bindin molecule in point blot assays, but did not show other proteins such as bovine serum albumin. This antibody cross-reactivity suggests that zosteric acid and bindin share significant structural similarity at the antibody recognition site, which is likely to be a sulfate moiety. Given the structural similarity at the sulfate moiety between bindin and zosteric acid, the question explains why zosteric acid is an effective inhibitor of sea urchin fertilization. Will.

Example 7: 4t-Pentylphenylchlorosulfate (4-PPCS) blocks fertilization The effect of 4-PPCS in inhibiting sea urchin fertilization, generally as described for zosteric acid in Example 6 went. As can be seen in FIG.
S was almost 100% effective at blocking sea urchin fertilization in the range of 1 to 10 mM, exactly the same range that zosteric acid was effective. As can be further seen in FIG. 7, when more sperm was added to the medium, 4-PPCS in solution
Beyond the binding capacity of PPCS, the inhibitory effects of PPCS may have been offset. In contrast to most biolethal agents, contact with 4-PPCS did not adversely affect sperm motility.

Equivalents Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

[Brief description of the drawings]

FIG. 1. The abundance of algal biofilm growth on inert coating RTV-11, RTV-11 when octyl sulfate was incorporated into the coating.
4 is a chart showing the results of a seaweed adhesion assay measured relative to the growth of the biofilm above. The relative algae abundance represents the adhesion of seaweed to the test surface.
Error bars indicate the standard deviation 1 of the median (n = 3) for each treatment. Wavelength 68 at time 0
The ratio of the optical densities measured at 0 nm and 750 nm (A ₆₈₀ / A ₇₅₀ ) was used as a baseline reference for all samples.

FIG. 2 is a chart showing the results of a bacterial adhesion assay performed by culturing marine bacterium Oceanosprilam in the presence and absence of either zosteric acid or methyl sulfate.

FIG. 3 is a chart showing the results of a bacterial adhesion assay performed by culturing marine bacterium Oceanosprilam in the presence and absence of either zosteric acid or octyl sulfate.

FIG. 4 is a chart showing the results of a bacterial adhesion assay performed using the bacterium Alteromonas-Atlantica in the presence and absence of either zosteric acid, octyl sulfate, or methyl sulfate.

FIG. 5 is a chart showing the results of a fungal attachment and growth assay using Aureobasidium-pullans (fungus in the raindrop that stains grout) grown in the presence and absence of zosteric acid. Sometimes the abundance of the fungus depends on the A. Indicates adhesion of pull lance.

FIG. 6: Measured in terms of percent transmission (% T) of the culture at a wavelength of 600 nm,
Figure 4 is a chart showing the results of bacterial Schwanella putritifaciens aggregation induced by the presence of increasing amounts of zosteric acid. Aggregation was indicated by a concentration-dependent increase in% T of the culture grown in the presence of zosteric acid. In this case, 8
The culture exposed to zosteric acid exhibited a relatively high level of% T due to the difference in growth since at time h, the count of viable colony forming units did not show any difference in cell density. It does not reflect that.

FIG. 7 shows the clotting time of red blood cells in the presence of zosteric acid,
FIG. 4 is a chart showing data from a prothrombin clotting time assay showing the clotting time of red blood cells in the presence of high molecular weight heparin.

FIG. 8 is a chart of data measuring the effect of zosteric acid on fertilization events in sea urchin eggs. a) shows the dose-dependent effect of zosteric acid on sea urchin fertilization. Percent fertilization compares the number of eggs fertilized in the presence of the indicated concentration of zosteric acid to the number of eggs fertilized under the same conditions in the absence of zosteric acid. b) shows the relative effect of equal concentrations (1 mg / mL) of coumaric acid, heparin and zosteric acid on the fertilization of sea urchin eggs.

FIG. 9: Various concentrations of 4t-pentylphenylchlorosulfate
It is a bar chart showing the fertilization of sea urchin eggs.

[Procedural Amendment] Submission of translation of Article 34 Amendment of the Patent Cooperation Treaty

[Submission date] September 29, 2000 (2000.9.29)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] Claims

[Correction method] Change

[Correction contents]

[Claims]

──────────────────────────────────────────────────続 き Continuation of front page (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE ), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SL, SZ, TZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AE, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN , IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, UA, UG, US, UZ, VN, YU, ZA, ZWF terms (reference) 4C206 AA01 AA02 JA06 MA01 MA04 NA14 ZB33 ZB35 ZC41

Claims (24)

[Claims] 1. A pharmaceutical carrier and an effective amount of the general formula 1: (Where X is -OH, -O (aryl), -O (acyl), -O (sulfonyl), -C
Represents N, F, Cl, or Br; Y represents O, S, Se, or NR; Z is an optionally substituted alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, Aralkyl, heteroaralkyl, or — (CH ₂ ) _m —R ₈₀ , where R is each independently hydrogen, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, or — (CH ₂ ) _m —R ₈₀ Wherein R ₈₀ each independently represents aryl, cycloalkyl, cycloalkenyl, heterocyclyl or polycyclyl, and m is an integer in the range from 0 to 8, including 8. Or a salt thereof. 2. The pharmaceutical composition according to claim 1, wherein X represents -OH, F, Cl, or Br. 3. The pharmaceutical composition according to claim 1, wherein Y represents O. During wherein wherein alkyl Z is substituted according to the selection, aryl, or _{- (CH} 2) represents an m _{-R 80,} A pharmaceutical composition according to claim 1. 5. The pharmaceutical composition according to claim 1, wherein Z represents an optionally substituted alkylphenyl, heteroalkylphenyl, arylphenyl, or heteroarylphenyl. 6. In the formula, Z is methyl, octyl, 4- (2-methylpropyl) phenyl, 4- (1,1-dimethylethyl) phenyl, 4- (1,1-dimethylpropyl) phenyl,
The pharmaceutical composition according to claim 1, which represents 4-pentylphenyl, 4- (1-methyl-1-phenylethyl) phenyl, or 4- (1-methylheptyl) phenyl. 7. The pharmaceutical composition according to claim 1, wherein R represents H or alkyl. 8. wherein X represents —OH, F, Cl, or Br; and Y represents O
The pharmaceutical composition according to claim 1, which represents: 9. The pharmaceutical composition according to claim 1, wherein X represents —OH or Cl and Y represents O. During 10. Formula, X denotes -OH, F, Cl, or Br, and alkyl Z is substituted according to the selection, aryl, or _- represents a _(CH 2) m _{-R 80,} wherein Item 7. The pharmaceutical composition according to Item 1. 11. The method of claim 1, wherein X represents —OH or Cl, and Z represents optionally substituted alkyl, aryl, or — (CH ₂ ) _m —R _80.
3. The pharmaceutical composition according to item 1. 12. The method of claim 12, wherein X represents —OH, F, Cl, or Br, and Z represents an optionally substituted alkylphenyl, heteroalkylphenyl, arylphenyl, or heteroarylphenyl. Item 7. The pharmaceutical composition according to Item 1. 13. The method of claim 1, wherein X represents —OH or Cl, and Z represents an optionally substituted alkylphenyl, heteroalkylphenyl, arylphenyl, or heteroarylphenyl. Pharmaceutical composition. 14. wherein X represents -OH, F, Cl or Br, and Z is methyl, octyl, 4- (2-methylpropyl) phenyl, 4- (1,1-dimethylethyl)
2. Representing phenyl, 4- (1,1-dimethylpropyl) phenyl, 4-pentylphenyl, 4- (1-methyl-1-phenylethyl) phenyl or 4- (1-methylheptyl) phenyl A pharmaceutical composition as described. 15. wherein X represents -OH or Cl, and Z is methyl, octyl, 4- (2-methylpropyl) phenyl, 4- (1,1-dimethylethyl) phenyl,
2. The method according to claim 1, which represents 4- (1,1-dimethylpropyl) phenyl, 4-pentylphenyl, 4- (1-methyl-1-phenylethyl) phenyl, or 4- (1-methylheptyl) phenyl. Pharmaceutical composition. During 16. wherein alkyl Y represents O, and wherein Z is substituted according to the selection, aryl, or _{- (CH} 2) represents an m _{-R 80,} A pharmaceutical composition according to claim 1 . 17. The pharmaceutical composition according to claim 1, wherein Y represents O and Z represents optionally substituted alkylphenyl, heteroalkylphenyl, arylphenyl, or heteroarylphenyl. . 18. A compound according to claim 18, wherein Y represents O and Z is methyl, octyl, 4- (2-
Methylpropyl) phenyl, 4- (1,1-dimethylethyl) phenyl, 4- (1,1-dimethylpropyl) phenyl, 4-pentylphenyl, 4- (1-methyl-1-phenylethyl)
The pharmaceutical composition according to claim 1, which represents phenyl or 4- (1-methylheptyl) phenyl. 19. In the formula, X represents —OH, F, Cl, or Br, and Y represents O.
And Z is optionally substituted alkyl, aryl, or-(CH₂) _m -R₈₀The pharmaceutical composition according to claim 1, which represents 20. wherein X represents -OH or Cl, Y represents O, and Z
Alkyl There substituted in accordance with the selection, aryl, or _{- (CH} 2) represents an m _{-R 80,} A pharmaceutical composition according to claim 1. 21. wherein X represents -OH, F, Cl, or Br, Y represents O, and Z is an optionally substituted alkylphenyl, heteroalkylphenyl, arylphenyl, or heteroaryl. 2. The pharmaceutical composition according to claim 1, which represents an arylphenyl. 22. wherein X represents —OH or Cl, Y represents O, and Z
The pharmaceutical composition according to claim 1, wherein represents an optionally substituted alkylphenyl, heteroalkylphenyl, arylphenyl, or heteroarylphenyl. 23. wherein X represents —OH, F, Cl, or Br, Y represents O, and Z is methyl, octyl, 4- (2-methylpropyl) phenyl, 4- (1, Represents 1-dimethylethyl) phenyl, 4- (1,1-dimethylpropyl) phenyl, 4-pentylphenyl, 4- (1-methyl-1-phenylethyl) phenyl, or 4- (1-methylheptyl) phenyl The pharmaceutical composition according to claim 1. 24. wherein X represents —OH or Cl, Y represents O, and Z
Is methyl, octyl, 4- (2-methylpropyl) phenyl, 4- (1,1-dimethylethyl) phenyl, 4- (1,1-dimethylpropyl) phenyl, 4-pentylphenyl, 4- (1-
The pharmaceutical composition according to claim 1, which represents methyl-1-phenylethyl) phenyl or 4- (1-methylheptyl) phenyl.